Kumar Sekar Vasantha, Srinath Selladurai, Saha Ranendra N
Birla Institute of Technology and Science (BITS), Formulation Development and Pharmacokinetic Laboratory, Pharmacy Group, Pilani, Rajasthan, India.
J Chromatogr Sci. 2012 Mar;50(3):259-70. doi: 10.1093/chromsci/bmr054.
A simple, highly repeatable and reproducible method for the estimation of Paclitaxel (TAX) in micro volume amounts of rat plasma is successfully developed and validated. The extraction procedure using 800 µL of ice-cold acetonitrile is very simple and economical with high sensitivity. The rectangular ratiograms and purity curve demonstrate the selectivity of the method. The validation and stability results show that propylparaben (PP) is a suitable internal standard (resolution 7.70 ± 0.15 min) for the estimation of TAX in micro volume rat plasma. TAX and PP are separated by isocratic reversed-phase high-performance liquid chromatography with diode array UV method with a retention time of 8.0 ± 0.25 and 5.3 ± 0.15 min, respectively, with a total run time of 10 min. The system suitability results show that the method has good reproducibility. The stability of TAX is well studied in rat plasma, and the % RSD of all stability studies of TAX are well within the acceptable range of ± 20 % at the lower limit of quantitation (LLOQ) and ± 15% at all quality control levels. The limit of detection (LOD) and LLOQ of the method are 5 and 10 ng/mL, respectively. This rapid method is successfully used to study the i.v pharmacokinetic of TAX at 10 mg/kg in wistar rats, and drug concentration is detected up to 24 h.
已成功开发并验证了一种用于测定微量大鼠血浆中紫杉醇（TAX）的简单、高度可重复且可再现的方法。使用800 μL冰冷乙腈的提取程序非常简单且经济，灵敏度高。矩形比例图和纯度曲线证明了该方法的选择性。验证和稳定性结果表明，对羟基苯甲酸丙酯（PP）是测定微量大鼠血浆中TAX的合适内标（分离度为7.70±0.15分钟）。TAX和PP通过等度反相高效液相色谱-二极管阵列紫外法分离，保留时间分别为8.0±0.25分钟和5.3±0.15分钟，总运行时间为10分钟。系统适用性结果表明该方法具有良好的重现性。已对TAX在大鼠血浆中的稳定性进行了充分研究，TAX所有稳定性研究的相对标准偏差（%RSD）在定量下限（LLOQ）时均在±20%的可接受范围内，在所有质量控制水平下均在±15%以内。该方法的检测限（LOD）和LLOQ分别为5和10 ng/mL。这种快速方法已成功用于研究10 mg/kg的TAX在Wistar大鼠体内的静脉药代动力学，药物浓度可检测长达24小时。
